Objective To dynamically observe changes of IgG, its subclasses and IgE in sera of mice by immunization with mixed recombinant of BCG-Em Ⅱ/3 and BCG-Em14-3-3 vaccine of Echinococcus multilocularis (Era). Methods Forty Balb/c mice of 12-14 week old and 20-25 g weight were intranasally vaccinated by the vaccine, 4 mice were killed randomly by the weight on 0,2,4,6,8,10,12,14,16 and 18 weeks of immunization respectively, sera were gathered from the eyeball to measure IgG, its subclasses and IgE by routine ELISA. Results Levels of IgG, IgG2a and IgG2b in the sera of mice increased obviously on 2-18 weeks, reached the highest level on 10, 4 and 4 weeks respectively, the value was 0.095±0.033,0.022±0.001,0.023±0.003 respectively, as compared with the value on 0 week(0.030±0.013,0.012±0.004,0.013±0.004), the difference being statistically significant(q=2.95,4.87,2.81 respectively, P < 0.01 or P < 0.05); levels of IgG1, IgG3 and IgE in the sera of mice decreased remarkably on 2-18 weeks,came to the lowest level on 4,2,6 weeks respectively, the value was 0.031±0.004,0.136±0.002,0.114±0.002 respectively, as compared with the value on 0 week(0.192±0.007, 0.175±0.013,0.024±0.003), the difference being statistically significant (q =5.16,4.93,5.32 respectively, P < 0.01 or P < 0.05). Conclusion Helper T cell(TH) Ⅰ response is induced in mice by mixed recombinant of BCG-Em Ⅱ/3 and BCG-Em14-3-3 vaccine on early immunization.

Abstract: Objective To explore the correlation between lung function in patients with chronic obstructive pulmonary disease (COPD) and tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) levels in exhaled breath condensate (EBC), and to provide a convenient methodological basis for the diagnosis and treatment of COPD and the determination of its efficacy. Methods A total of 81 COPD patients and 40 healthy controls were selected from the respiratory department of the Fourth Affiliated Hospital of Guangzhou Medical University from August 2020 to February 2022 as the research subjects. The COPD patients were divided into 41 cases in the acute exacerbation group and 40 cases in the remission group according to their status. All participants underwent lung function detection, venous blood and EBC collection, and the levels of TNF-α and IL-1β in EBC and venous blood were analyzed by enzyme-linked immunosorbent assay (ELISA), and correlation analysis was performed by Pearson correlation analysis method. Results The levels of TNF-α and IL-1β in EBC of in the acute exacerbation group, the healthy control group, the remission group were (5.16±0.18) pg/μL and (7.75±0.27) pg/μL, (2.66±0.31) pg/μL and (2.41±0.24） pg/μL, (3.61±0.29) pg/μL and (3.17±0.38) pg/μL, respectively. Compared with the healthy control group, the levels of TNF-α and IL-1β in EBC in the COPD acute exacerbation group were significantly higher than those in the healthy control group and the COPD remission group (F=9.451, 8.217, P<0.001). Serum tests were consistent with this result. Correlation analysis showed that the levels of TNF-α and IL-1β in EBC were significantly positively correlated with the level of serum inflammation levels (P<0.001), while significantly negatively correlated with lung function (P<0.001).  Conclusions TNF-α and IL-1β in EBC are potential biomarkers of inflammation in patients with COPD, and their detection can be used to effectively assess lung function in patients with COPD. 

ObjectiveTo explore the renal functions of vasopressin-deficient Brattleboro(BB) rats by using different concentrated Gadolinium-diethylenetetramine pentaacetic acid(Gd-DTPA) dynamic enhanced magnetic resonance imaging(MRI).MethodsThe study included 14 BB rats(male rats of 3 month-old) and 14 normal male Wistar rats used as control group.Dynamic MRI was performed by using either a low dosage(0.05 mmol/kg) or a high dosage of Gd-DTPA(0.5 mmol/kg).Data of 0-60 min renal cortex,medulla and pelvic were obtained after using contrast medium.MRI of kidneys at different time was analyzed and the mean relative signal intensity(RSI) was measured.Then the RSI curves of different groups were marked.Data of each group were caculated separately by SPSS 11.0 software.ResultsThe findings demonstrated that RSI curves of the vasopressin-deficient kidneys showed different patterns as compared with those of the control group(P

Objective To optimize methods of culturing smooth muscle progenitor cells(SPCs) from mononuclear cells(MNCs) of peripheral blood. Methods Human MNCs isolated from buffy coat were seeded on M-199 with bovine pituitary extraction.On the eighth day outgrowth cells were stimulated with platelet-derived growth factor-BB(PDGF-BB).Fifteen days later,immunofluorescence,Western blot or RT-PCR was used to analyzed the expression of smooth muscle cell specific ?-actin(?-SMA),smooth muscle myosin heavy chain(SM MHC),Calponin,CD34,Tie-2 and Flk-1,and fluorescence activated cell sorter was employed to examine ?-SMA positive cells ratio. Results The cells stimulated by PDGF-BB for 15 d were positive for ?-SMA,SM MHC,Calponin,CD34 and Flk-1,but negative for Tie-2.The ?-SMA positive cells ratio was(90.57?5.63)%,significantly different from that of the control(P

Objective To evaluate the clinical efficacy of endourologic treatment of benign uret-erointestinal anastomotic strictures in patients with urinary diversion. Methods Nine cases of benign ureterointestinal anastomotic strictures with a length of 1-3 cm following radical cystectomy and uri-nary diversion accepted endourologic treatment. 8 cases were treated by antegrade percutaneous ap-proach, 1 case by retrograde ureteroscopic approach. The strictures received balloon dilation, and ure-teral stents indewelled. Results In a follow up of 0.5-5.0 years, 1 case received percutaneous ne-phrostomy for complete ureterointestinal anastomotic atresia and refused to open operation reconstruc-tion. 5 cases had no recurrence after 2-3 endoscopic sessions. 3 cases needed long time ureteral stents indwelled. Conclusion Endourological technique for ureterointestinal strictures following urinary di-version avoided the disadvantages of open operation.

Objective To investigate the renal pelvic pressure(RPP) during minimally invasivepereutaneous nephrolithotomy(MPCNL),and inspect its influence to postoperative fever. MethodsThe RPP was measured by baroeeptor,and these data about pressure and postoperative fever wereevaluated statistically. Results The mean RPP was 14.72 mm Hg,the mean accumulative time of RPP≥30 mm Hg was 116.06 s. Fifteen cases(18. 75%)had a postoperative fever. Logistical analysissuggested that postoperative fever did not correlate to sex(P=0.195),age(P=0.641),urinary tractinfection (P=0.663),white blood cell≥10 × 109/L in blood routine examination postoperatively (P=0.751),once an occurrence of RPP≥40 mm Hg(P=0.662),while infection calculi (P=0.000),percutaneous tract size(P=0.029),mean RPP(P=0.036) ,mean RPP≥20 mm Hg(P=0.013),accumulative time of RPP≥30 mm Hg(P=0.010) and RPP≥30 mm Hg longer than 50 s(P=0.024)contributed to postoperative fever. Conclusions Renal pelvic pressure generally remains lower than alevel to back flow (30 mm Hg) during MPCNL. A transient renal pelvic pressure≥30 mm Hg don'tcountribute to postoperative fever,while a temporary high pressure status(50 s)would had an accumulated effect which means an enough back flow to bring a fever.

AIMTo study the possibility that responses of fever and discharge of neurons in PVN to intraperitoneal administration of LPS are mediated by vagal afferents.

METHODSRectal temperature of rat was detected by digital temperature detecting instrument. Glass micropipette placed in PVN was used to record unit discharges of neurons in it, before and after LPS was injected into PVN in normal rats and vagotomy rats.

RESULTSThe rectal temperature change value in vagotomy LPS group was significantly decreased compared with that in sham LPS group, and there was striking difference between them (P < 0.05). The discharges of neurons in PVN was increased in the normal rat in response to LPS. The discharges of neurons in PVN had no significant change in the vagotomy rats in response to LPS.

CONCLUSIONThe results indicate that vagus nerve may be one of the pathways of peripheral LPS signal communicating to CNS.

Animals ; Diaphragm ; innervation ; Fever ; chemically induced ; physiopathology ; Lipopolysaccharides ; Male ; Neurons ; physiology ; Paraventricular Hypothalamic Nucleus ; physiopathology ; Rats ; Rats, Wistar ; Vagotomy, Truncal

Objective To construct and identify recombinant Bifutobacteria (rBb)-Eg95 vaccine of Echinococcus granulosus (Eg). Methods The total RNA was extracted from hydatid cyst protoscoleces shattered by ultrasound, Eg95 antigen encoding gene was obtained by reverse transcription-polymerase chain reaction(RT-PCR) from the template of total RNA using the primer designed according to the DNA sequence of Eg95, the gene was cloned into Escherichia coli-Bifutobacteria(E.coli-Bb) shuttle plasmid pGEX-1λT and transformed into E.coli BL2 (DE3) competent cell to construct recombinant plasmid pGEX-Eg95 using BamH Ⅰ and EcoR Ⅰ, the recombinant plasmid was identified by restriction endonuclease digestion, then was electroporated into Bb to construct rBb-Eg95 vaccine, the vaccine was identified by PCR. Results Four hundred and seventy-one bp Eg95 gene was amplified by RT-PCR, the products of restriction endonuclease digestion were the same as expected(471 bp Eg95 gene and 4947 bp pGEX-1λT), 471 bp Eg95 gene fragment was amplified by PCR from the template of pGEX-Eg95 extracted from rBb vaccine. Conclusion rBb-Eg95 vaccine of Eg is successfully constructed, which lays the theoretical foundation for exploitation and utilization of this vaccine.

Objective To dynamically observe the changes of cytokines of splenocytes in mice immunized with recombinant bifidobacteria bifidum (Bb)- Eg95-EgA31 vaccine of Echinococcus grauulosus (Eg). Methods Balb/c mice were vaccinated by 5× 108 colony forming unit(CFU) orally and 5 × 105 CFU intranasally, respectively.Mice were killed on week 0,2,4,6,8,10, 12,14,16, 18 and 20 after immunization, respectively, and spleens were separated for cell culture with the stimulation of EgAg, concanavalin A (ConA) or lipopolysaccharide (LPS). The splenocyte supernatants were collected to determine the levels of interferonγ(IFN-γ), interleukin(IL)-12, tumor necrosis factor α(TNF-o) and IL-l0 using enzyme linked immunosorbent assay(ELISA) with MRS as control. Results In the oral immunization group, the levels of IFN-γ, IL-12, TNF-α and IL-10 showed a significant increase from week 2 to week 8, week 2 to week 8, week 4 and week 6 to week 10 after vaccination, respectively, and reached the highest level on week 4, week 2, week 4 and week 6 after vaccination, respectively;in EgAg stimulation group, the levels of IFN-γ, IL-12, TNF-α and IL-10 were (700.0 ± 115.5), (45.0 ± 5.8), (350.0 ± 57.7), (112.5 ± 14.4)ng/L, respectively, compared with week 0[(35.0 ± 5.8), (12.5 ± 2.9), (190.0 ± 11.6), (25.0 ± 5.8)ng/L, P ＜0.05 or ＜ 0.01] and MRS control group[(37.5 ± 5.0),(13.8 ± 2.5), (195.0 ± 5.8), (27.5 ± 2.9)ng/L, P＜ 0.05or ＜ 0.01]. In the intranasal immunization group, the levels of IFN-γ, IL-12, TNF-α and IL-10 showed an obvious increase from week 2 to week 8, week 2 to week 8, week 2 to week 6 and week 6 to week 16 after vaccination,respectively, and reached the highest level on week 2, week 2, week 4 and week 8 after vaccination, respectively;in EgAg stimulation group, the levels of IFN-γ, IL-12, TNF-α and IL-10 were (700.0 ± 115.5), (55.0 ± 5.8),(275.0 ± 28.9), (140.0 ± 11.6)ng/L, compared with week 0[(35.0 ± 5.8), (12.5 ± 2.9), (190.0 ± 11.6), (25.0 ±5.8)ng/L, P ＜ 0.05 or ＜ 0.01] and MRS control group[(37.5 ± 5.0), (13.8 ± 2.5), (195.0 ± 5.8), (27.5 ± 2.9)ng/L, P ＜ 0.05 or ＜ 0.01]. The cytokine levels in the groups with EgAg, ConA or LPS stimulus were significantly higher than those in the corresponding splenocytes suspension groups(P ＜ 0.05 or ＜ 0.01) , and the cytokine levels in the groups with ConA or LPS stimulus were obviously higher than those in the corresponding groups with EgAg stimulation(P ＜ 0.05 or ＜ 0.01). Conclusion The mixed Th1 and Th2 type response can be induced in mice immunized with the recombinant Bb-Eg95-EgA31 vaccine of Echinococcus granulosus in the early stage of immunization(2 to 6weeks).

Objective　To evaluate the effect of Naogongfude (NGFD, a traditional Chinese medicine orally administrated) on learning and memory and the expressions of synaptophysin (SY) and Tau-protein in cerebral cortex and hippocampus in aged rats and to explore the possible mechanism. Methods　Rats were divided into normal and neural disturbance groups according to the outcomes of active avoidance reaction (AAR) test, and then each group was randomly divided into control and NGFD-treated experimental groups. Animals were orally fed with NGFD for 2 months (5 ml/d) in experimental group or routinely fed in control and taken AAR and passive avoidance reaction (PAR) tests. After the rats were sacrificed, the synaptosome count, the expression of SY and Tau-protein, and the neuron apoptosis in cerebrum were examined. Results　The rats after 2-month NGFD administration had an increased AAR acquisition, obviously delayed AAR extinction and prolonged step through latency (STL) of PAR. The number of synaptosomes was raised and the immunoreactive intensity of synaptophysin was increased remarkedly, while Tau-protein immunoreactivity and apoptotic cells were decreased in cerebrum. Conclusion　NGFD does have the effect of improving brain function and putting off the aging of rat brain according to the results of behavior study and morphological observation.